牙髓炎症诱导P2X受体家族在外周及中枢神经系统中的表达
发布时间：2019-02-13 05:42
【摘要】：第一部分LPS诱导牙髓炎症过程中P2X受体在三叉神经节中的表达和作用 研究目的：离子通道型嘌呤受体家族(P2X)可以参与调节初级感觉神经元中的疼痛感觉。本实验目的在于研究在牙髓炎症引起三叉神经节(trigeminal ganglion,TG)中神经元活化以及P2X受体的表达模式,并采用P2X受体抑制剂观察抑制P2X受体后对痛觉传导相关因子表达的改变的影响。 研究方法：本实验将脂多糖(Lipopolysaccharides,LPS)置于大鼠左侧的上颌第一磨牙髓腔,以诱导牙髓炎症。观察了急性牙髓炎症期TG中活化神经元标志c-fos的分布,P2X受体与谷氨酸能神经元和GABA能神经元的关系,并采用免疫组化和蛋白印迹技术观察了TG的V1-V2支中的P2X受体的表达模式。进一步在髓腔中置入P2X受体抑制剂TNP-ATP并同时于大鼠左侧须垫注射TNP-ATP,观察了抑制P2X受体对急性牙髓炎症期TG中c-fos,谷氨酸能神经元标志VGluT1和GABA能神经元标志GAD67表达,以及p38和ERK1/2磷酸化的影响。 研究结果：FG逆行性标记显示同侧TG的V1-V2支支配左上颌第一磨牙,牙髓炎症可导致V1-V2支中神经元活化的标志c-fos表达增加；P2X2, P2X3和P2X5受体在TG的神经元胞浆中表达,在牙髓炎症进展的第1天表达增加,至第三天持续增加；P2X2, P2X3和P2X5受体在V1-V2支中的表达增高主要发生在与痛觉传导相关的小直径神经元中。荧光双标记法显示P2X2,P2X3和P2X5受体在V1-V2支中分别与VGluT1和GAD67共表达在小直径的TG神经元的胞浆中；LPS刺激牙髓诱导炎症能导致c-fos, VGluT1和GAD67表达增高,并导致p-p38和p-ERK1/2表达增加；抑制P2X受体会导致c-fos, VGluT1和GAD67的表达增高受到抑制,并降低p-p38和p-ERK1/2表达。 研究结论：LPS诱导的急性牙髓炎症可诱导P2X2, P2X3和P2X5受体在活化的TGV1-V2支中的表达增高,提示P2X受体可能参与了牙髓炎症诱导的神经敏感性增高。抑制P2X受体会使牙髓炎症引起的TG中c-fos,VGluT1和GAD67的表达增高受到抑制,并降低p38和ERK1/2的磷酸化。 第二部分牙髓炎症对大鼠中枢神经系统中P2X表达的影响 研究目的：本实验拟采用大鼠开髓模型诱导大鼠牙髓炎症,目的在于研究不同牙髓炎症进展阶段对脑干的三叉神经脊束核尾端亚核(Vc)和丘脑的腹后中核(VPM)中的影响。 研究方法：本实验在大鼠左上颌第一磨牙颌面开髓,并暴露与口腔菌群环境,诱导牙髓炎症。取牙髓炎症第1天,第3天,第7天和第28天时,大鼠双侧脑干的三叉神经脊束核尾端亚核(Vc)和丘脑的腹后中核(VPM),采用免疫组化和免疫荧光方法观察P2X受体家族(P2X1-7受体)的表达模式。 研究结果：实验观察到牙髓炎症第1天,第3天,第7天显示急性牙髓炎症的组织学表现,其中第3天最为典型,而第28天显示牙髓坏死和慢性根尖周炎的组织学表现；P2X1-6受体在Vc和VPM中均表达在神经元的胞浆中,而P2X7表达在Vc和VPM中在小胶质细胞中；P2X2,P2X4和P2X7受体在急性牙髓炎症期在同侧Vc中的表达增加；P2X1-5和P2X7的表达都在双侧VPM中均上升,开髓3d时,对侧VPM中P2X1-5和P2X7的平均表达面积显著高于同侧。 研究结论：除P2X6受体持续高表达外,P2X1-5和P2X7受体均可受到牙髓炎症的诱导,在Vc和VPM中表达增高。
[Abstract]:The expression and function of P2X receptor in the trigeminal ganglion during the first part of LPS-induced dental pulp inflammation The purpose of this study is that the ion channel type methotrexate receptor family (P2X) can be involved in the regulation of the pain in primary sensory neurons. The purpose of this study was to study the activation of the neurons in the trigeminal ganglion (TG) and the expression pattern of the P2X receptor in the pulpal inflammation, and to use the P2X receptor inhibitor to observe the effect of the P2X receptor on the expression of the hyperalgesia-related factors. In this experiment, Lipopolysaccharide (LPS) was placed on the left side of the rat's maxillary first pulp cavity to induce the teeth. The distribution of c-fos, the distribution of the c-fos, the relationship between the P2X receptor and the neuronal and GABA-ergic neurons in the TG of acute endodontic inflammation were observed, and the expression of the P2X receptor in the V1-V2 branch of TG was observed by immunohistochemistry and Western blotting. In the model, the P2X receptor inhibitor TNP-ATP was further placed in the medullary cavity and the TNP-ATP was injected at the left side of the rat, and the expression of the c-fos, the glutamate-capable neuronal marker VGluT1 and the GABA-enabled neuronal marker GAD67 in the acute endodontic inflammation phase TG, and the phosphorylation of p38 and ERK1/ 2 were observed. The results of the study: The reverse marker of FG showed that the V1-V2 branch of the same side was dominant to the left maxillary first molar, which could lead to the increase of the expression of c-fos in the neurons in the V1-V2 branch, and the expression of the P2X2, P2X3 and P2X5 receptors in the cytoplasm of the neurons in the TG, and the first in the progress of the dental pulp inflammation. The increase of the expression of P2X2, P2X3 and P2X5 in the V1-V2 branch mainly occurred in the small straight line related to the conduction of the hyperalgesia. The expression of P2X2, P2X3 and P2X5 was co-expressed with VGluT1 and GAD67 in the cytoplasm of the small-diameter TG neurons, and the expression of p-p38 and p-ERK1/ 2 increased and the expression of p-p38 and p-ERK1/ 2 increased, and the inhibition of P2X receptors could lead to increased expression of p-p38 and p-ERK1/ 2. The expression of c-fos, VGluT1 and GAD67 was inhibited and the p-p38 and p-ERK were reduced. Conclusion: The expression of P2X2, P2X3 and P2X5 receptor in the activated TGV1-V2 branch can be induced by LPS-induced acute dental pulp inflammation, suggesting that the P2X receptor may be involved in the induction of dental pulp inflammation. The inhibition of the expression of c-fos, VGluT1 and GAD67 in the TG induced by the pulpal inflammation by the P2X receptor is inhibited and p38 and ERK are reduced. 1/ 2 phosphorylation. The second part of dental pulp inflammation is the central nervous system of the rat The effect of P2X expression in the system is the purpose of the study: this experiment is to be used in rats The purpose of this study was to study the subnucleus (Vc) of the nucleus of the trigeminal nerve of the brain stem and the ventral part of the thalamus in the stage of different pulp inflammation. The effect of this experiment on the maxillary first molar and maxillofacial opening of the rat's left maxillary first molar and the exposure to The first day, third day, day 7 and day 28 of the dental pulp inflammation, the subnucleus (Vc) and the nucleus (VPM) of the posterior subnucleus (VPM) of the nucleus of the trigeminal nerve of the rat's dual-side brain stem were observed, and the P2X receptor family (P) was observed by the method of immunohistochemistry and immunofluorescence. The expression pattern of the 2X1-7 receptor was observed. The results of the study showed that the first day, the third day and the seventh day of the dental pulp inflammation showed the histological appearance of the acute dental pulp inflammation, in which the third day was the most typical, and the dental pulp was shown on the 28th day. The expression of P2X2, P2X4, and P2X7 in the same side of Vc and VPM increased with the expression of P2X2, P2X4, and P2X7 in the same side, and the expression of P2X1-5 and P2X7 was in the same side. Both sides VPM rise and open the nucleus for 3d, P2X1-5 and P2 in the side VPM The average expression area of X7 was significantly higher than that of the same side. Conclusion: In addition to the high expression of P2X6 receptor, P2X1-5 and P2X7 receptors can be affected by pulpitis.
【学位授予单位】：武汉大学
【学位级别】：博士
【学位授予年份】：2014
【分类号】：R781.31
本文编号：2421235
[Abstract]:The expression and function of P2X receptor in the trigeminal ganglion during the first part of LPS-induced dental pulp inflammation The purpose of this study is that the ion channel type methotrexate receptor family (P2X) can be involved in the regulation of the pain in primary sensory neurons. The purpose of this study was to study the activation of the neurons in the trigeminal ganglion (TG) and the expression pattern of the P2X receptor in the pulpal inflammation, and to use the P2X receptor inhibitor to observe the effect of the P2X receptor on the expression of the hyperalgesia-related factors. In this experiment, Lipopolysaccharide (LPS) was placed on the left side of the rat's maxillary first pulp cavity to induce the teeth. The distribution of c-fos, the distribution of the c-fos, the relationship between the P2X receptor and the neuronal and GABA-ergic neurons in the TG of acute endodontic inflammation were observed, and the expression of the P2X receptor in the V1-V2 branch of TG was observed by immunohistochemistry and Western blotting. In the model, the P2X receptor inhibitor TNP-ATP was further placed in the medullary cavity and the TNP-ATP was injected at the left side of the rat, and the expression of the c-fos, the glutamate-capable neuronal marker VGluT1 and the GABA-enabled neuronal marker GAD67 in the acute endodontic inflammation phase TG, and the phosphorylation of p38 and ERK1/ 2 were observed. The results of the study: The reverse marker of FG showed that the V1-V2 branch of the same side was dominant to the left maxillary first molar, which could lead to the increase of the expression of c-fos in the neurons in the V1-V2 branch, and the expression of the P2X2, P2X3 and P2X5 receptors in the cytoplasm of the neurons in the TG, and the first in the progress of the dental pulp inflammation. The increase of the expression of P2X2, P2X3 and P2X5 in the V1-V2 branch mainly occurred in the small straight line related to the conduction of the hyperalgesia. The expression of P2X2, P2X3 and P2X5 was co-expressed with VGluT1 and GAD67 in the cytoplasm of the small-diameter TG neurons, and the expression of p-p38 and p-ERK1/ 2 increased and the expression of p-p38 and p-ERK1/ 2 increased, and the inhibition of P2X receptors could lead to increased expression of p-p38 and p-ERK1/ 2. The expression of c-fos, VGluT1 and GAD67 was inhibited and the p-p38 and p-ERK were reduced. Conclusion: The expression of P2X2, P2X3 and P2X5 receptor in the activated TGV1-V2 branch can be induced by LPS-induced acute dental pulp inflammation, suggesting that the P2X receptor may be involved in the induction of dental pulp inflammation. The inhibition of the expression of c-fos, VGluT1 and GAD67 in the TG induced by the pulpal inflammation by the P2X receptor is inhibited and p38 and ERK are reduced. 1/ 2 phosphorylation. The second part of dental pulp inflammation is the central nervous system of the rat The effect of P2X expression in the system is the purpose of the study: this experiment is to be used in rats The purpose of this study was to study the subnucleus (Vc) of the nucleus of the trigeminal nerve of the brain stem and the ventral part of the thalamus in the stage of different pulp inflammation. The effect of this experiment on the maxillary first molar and maxillofacial opening of the rat's left maxillary first molar and the exposure to The first day, third day, day 7 and day 28 of the dental pulp inflammation, the subnucleus (Vc) and the nucleus (VPM) of the posterior subnucleus (VPM) of the nucleus of the trigeminal nerve of the rat's dual-side brain stem were observed, and the P2X receptor family (P) was observed by the method of immunohistochemistry and immunofluorescence. The expression pattern of the 2X1-7 receptor was observed. The results of the study showed that the first day, the third day and the seventh day of the dental pulp inflammation showed the histological appearance of the acute dental pulp inflammation, in which the third day was the most typical, and the dental pulp was shown on the 28th day. The expression of P2X2, P2X4, and P2X7 in the same side of Vc and VPM increased with the expression of P2X2, P2X4, and P2X7 in the same side, and the expression of P2X1-5 and P2X7 was in the same side. Both sides VPM rise and open the nucleus for 3d, P2X1-5 and P2 in the side VPM The average expression area of X7 was significantly higher than that of the same side. Conclusion: In addition to the high expression of P2X6 receptor, P2X1-5 and P2X7 receptors can be affected by pulpitis.
【学位授予单位】：武汉大学
【学位级别】：博士
【学位授予年份】：2014
【分类号】：R781.31
【参考文献】
相关期刊论文 前1条
1 刘宇炜;陈晓青;田香;陈琳;吴钰祥;黄丹;易卉玲;易初丽;李超英;;P2X_3 , but not P2X_1 , Receptors Mediate ATP-activated Current in Neurons Innervating Tooth-pulp[J];Journal of Huazhong University of Science and Technology(Medical Sciences);2013年03期
，本文编号：2421235
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